Highly soluble stevia sweetener

ABSTRACT

A method for making highly soluble  Stevia  sweetener compositions is described. The resulting sweetener compositions readily provide high concentration solutions, and also possess superior taste qualities. The compositions can be used as sweeteners, sweetness enhancers, and flavor enhancers in foods, beverages, cosmetics and pharmaceuticals.

FIELD OF THE INVENTION

The invention relates to a process for the preparation of highly solublesweet glycosides from a Stevia rebaudiana Bertoni plant, and moreparticularly for preparation of highly soluble compositions containingrebaudioside D.

DESCRIPTION OF THE RELATED ART

The extract of Stevia rebaudiana plant (Stevia) contains a mixture ofdifferent sweet diterpene glycosides, which have a commonaglycon—steviol and differ by the presence of carbohydrate residues atpositions C13 and C19 of steviol molecule. These glycosides accumulatein Stevia leaves and compose approximately 10%-20% of the total dryweight. Typically, on a dry weight basis, the four major glycosidesfound in the leaves of Stevia are Dulcoside A (0.3%), Rebaudioside C(0.6-1.0%), Rebaudioside A (3.8%) and Stevioside (9.1%). Otherglycosides identified in Stevia extract include Rebaudioside B, C, D, E,and F, Steviolbioside and Rubusoside. Steviol glycosides differ fromeach other by their taste properties. Some of them possess significantbitterness, lingering licorice aftertaste (Prakash I., DuBois G. E.,Clos J. F., Wilkens K. L., Fosdick L. E. (2008) Development of rebiana,a natural, non-caloric sweetener, Food Chem. Toxicol., 46, S75-S82).

Rebaudioside D or Reb D (CAS No: 63279-13-0), is one of the sweetglycosides found in Stevia rebaudiana. Studies show that Reb D possessvery desirable taste profile, almost lacking bitterness, and lingeringlicorice aftertaste typical for other Steviol glycosides.

These properties multiply the significance of Reb D and attract greatinterest for methods of preparation of Reb D.

However highly purified steviol glycosides possess relatively low watersolubility. For example Rebaudioside A (Reb A) thermodynamic equilibriumsolubility at room temperature is only 0.8% .

On the other hand, it is well known that steviol glycosides exhibitso-called polymorphism (Zell T. M., Padden B. E., Grant D. J. W.,Schroeder S. A., Wachholder K. L., Prakash I., Munsona E. J. (2000)Investigation of Polymorphism in Aspartame and Neotame Using Solid-StateNMR Spectroscopy, Tetrahedron, 56, 6603-6616). Particularly Reb Aamorphous, anhydrous and solvate forms differ significantly from eachother in terms of solubility which is one of the main criteria for thecommercial viability of a sweetener. In this regard, as shown in Table1, the hydrate form of Reb A displays the lowest solubility (Prakash I.,DuBois G. E., Clos J. F., Wilkens K. L., Fosdick L. E. (2008)Development of rebiana, a natural, non-caloric sweetener, Food Chem.Toxicol., 46, S75-S82). It was shown that Reb A may transform from onepolymorph form to another at certain conditions (U.S. patent applicationSer. No. 11/556,049).

TABLE 1 Properties of Rebaudioside A forms (US Pat. Appl. 11/556,049)Polymorph Forms Form 1 Form 2 Form 3 Form 4 Hydrate Anhydrous SolvateAmorphous Rate of Very low Intermediate High (>30% High (>35%dissolution in (<0.2% in (<30% in 5 in 5 minutes) in 5 minutes) H₂O at25° C. 60 minutes) minutes) Alcohol <0.5% <1% 1-3% <0.05% contentMoisture   >5% <1%  <3%  6.74% content

Reb D possesses even lower water solubility compared to Reb A. At roomtemperature it can be dissolved only at 0.01-0.05%. When heat is appliedone can make up to 0.5% solution. However, upon cooling to roomtemperature, Reb D will quickly crystallize back from solution.Considering high sweetness intensity of Reb D (about 200 times of sugarsweetness)—even 0.05% solubility may seem sufficient for manyapplications.

Many food production processes use highly concentrated ingredient mixesprior to producing final forms of food products. In that case, muchhigher concentrations of dissolved Reb D will be required. It is notedthat using heat for dissolution of Reb D may not be possible incompositions containing heat sensitive components. Also maintaining hightemperature of mixture for prolonged time to prevent prematurecrystallization of Reb D can cause thermal degradation of mixturecomponents or undesirable changes of organoleptic properties.

U.S. patent application Ser. No. 12/612,374 describes a preparationmethod of anhydrous form of Reb D which possesses about 0.15% solubilityat 50° C. The method requires heat treatment of Reb D powder at atemperature 80°-110° C. for about 50-120 hours, under vacuum. It isnoted that the described method allows preparing Reb D form that haslimited solubility and still requires significant increase oftemperature for dissolution. Extended time period of said vacuum thermaltreatment is a disadvantage as well.

U.S. patent application Ser. No. 12/700,223 describes method ofpreparing supersaturated solutions of Reb D, wherein the mixture of RebD with aqueous liquid is heated to 75°-90° C. and then gradually cooledto provide relatively stable 0.3% Reb D solution. It is noted that thesaid method describes only a solubilization technique for a specificaqueous liquid, and does not provide readily usable form of highlysoluble Reb D.

U.S. patent application Ser. No. 13/022,727 describes inclusioncomplexes of steviol glycosides and cyclodextrins, wherein glycoside tocyclodextrin ratio ranges from 1:1 to 1:20 and the solubility of saidcomplexes range from 0.1-7%. It is noted that some techniques describedwithin the application, such as freeze-drying, are difficult toimplement in large-scale multi-ton productions. The significant portionon non-sweet (low sweetness) compound, (i.e. cyclodextrin), reduces theoverall sweetness of the mixture as well.

There is a need for developing highly soluble forms or compositions ofReb D that possess high sweetness intensity, and provide stablesolutions at room temperature.

Furthermore, considering the similar chemical structures of Reb D andother steviol glycosides, as well as other terpene glycosides, there isalso a need to develop approaches that may be used in case of otherglycosides as well.

SUMMARY OF THE INVENTION

The invention is directed to a method for producing a sweetenercomprising the steps of providing Stevia sweetener powder, solubilizingit in water under gradient temperature treatment conditions, to producehighly stable concentrated solution, and spray drying the highly stableconcentrated solution to obtain a highly soluble Stevia sweetenerpowder.

Hereinafter the term “steviol glycoside(s)” will mean Rebaudioside A,Rebaudioside B, Rebaudioside C, Rebaudioside D, Rebaudioside E,Rebaudioside F, Stevioside, Steviolbioside, Dulcoside A, Rubusoside, orother glycoside of steviol and combinations thereof.

Hereinafter, unless specified otherwise the solubility of material isdetermined in reverse osmosis (RO) water at room temperature. Where thesolubility is expressed as “%” it to be understood as number of grams ofmaterial soluble in 100 grams of solvent.

It is to be understood that both the foregoing general description andthe following detailed description are exemplary and explanatory and areintended to provide further explanation of the invention as claimed.

DETAILED DESCRIPTION OF THE INVENTION

A process for the preparation of highly soluble Stevia sweetener,particularly a Stevia sweetener comprising Reb D, is described herein.

Crystalline Reb D has an inherently low solubility, ranging from about0.01%-0.05%. The need exists, therefore, for a process in which a highsolubility Reb D or compositions thereof are prepared.

In one embodiment of the present invention, initial materials,comprising sweet glycoside(s) of the Stevia rebaudiana Bertoni plantextract, which includes Rebaudioside D, Stevioside, Rebaudioside A,Rebaudioside B, Rebaudioside C, Rebaudioside E, Rebaudioside F,Steviolbioside, Dulcoside A, Rubusoside or other glycoside of stevioland combinations thereof were combined with the water at ratio of 1:1 to1:10, preferably 1:3 to 1:6 (w/v).

The obtained mixture was further subjected to a gradient heat treatmentthat resulted in high stability and high concentration solution. Thegradient of 1° C. per minute was used in heating the mixture. Themixture was heated to the temperature of 110-140° C., preferably118-125° C. and was held at maximum temperature for 0-120 min,preferably 50-70 min.

After the heat treatment the solution was cooled down to about 80° C. atgradient of 1° C. per minute. This high stability and high concentrationsolution did not show any crystallization during up to 1-hourincubation.

The solution was spray dried by laboratory spray drier operating at 175°C. inlet and 100° C. outlet temperature. A highly soluble amorphouscompositions of Reb D were obtained with >1% solubility in water at roomtemperature.

In another embodiment of this invention the initial materials wereselected from the group including Reb D, Reb A, Rebaudioside B (Reb B),and steviolbioside (Sbio)

In yet another embodiment combining the Reb D and Reb B, treating andspray drying it by the methods described above, yields a compositionwith significantly higher solubility (about 1%) compared to thecomposition obtained by combination of Reb D and Reb A at the same ratio(about 0.5%). This phenomenon was unexpected, as both Reb D and Reb Bhave <0.1% solubility, whereas the Reb A used in the experiment had >5%solubility. Therefore combinations of Reb D and Reb A were expected tohave higher solubility compared to combinations of Reb D and Reb B.

In one embodiment the said combination comprises Reb D and Reb B at aratio from 50%:50% to 80%:20%, preferably 70%:30% to 80%:20%. Theresulting product has a solubility ranging from 0.5% to 2%.

In yet another embodiment the Reb B was fully or partially, convertedinto carboxylate salt form. The solution containing RebD and Reb B,after heat treatment and subsequent cooling (as described above), wasmixed with solution of base to achieve a pH level of 4.5-7.0, preferablypH 5.5-6.5. The obtained mixture was spray dried as described above.Alternatively other reactions able to convert Reb B into carboxylatesalt form may be used as well. The preferred cations were K⁺ and Na⁺,and the bases—KOH and NaOH respectively. However those skilled in artshould recognize that other carboxylate salts of Reb B can be preparedin a similar manner.

In one embodiment the taste properties of compositions comprising Reb Dand Reb B were compared with compositions comprising Reb D and Reb A atthe same ratio. The evaluation shows that combinations of Reb D and RebB possessed superior taste properties compared to combinations of Reb Dand Reb A. Particularly the compositions of Reb D and Reb B showedalmost no lingering and bitterness, and more rounded sugar-like tasteprofile.

The obtained compositions can be used as sweetener, sweetness enhancer,and flavor enhancer in various food and beverage products. Non-limitingexamples of food and beverage products include carbonated soft drinks,ready to drink beverages, energy drinks, isotonic drinks, low-caloriedrinks, zero-calorie drinks, sports drinks, teas, fruit and vegetablejuices, juice drinks, dairy drinks, yoghurt drinks, alcohol beverages,powdered beverages, bakery products, cookies, biscuits, baking mixes,cereals, confectioneries, candies, toffees, chewing gum, dairy products,flavored milk, yoghurts, flavored yoghurts, cultured milk, soy sauce andother soy base products, salad dressings, mayonnaise, vinegar,frozen-desserts, meat products, fish-meat products, bottled and cannedfoods, tabletop sweeteners, fruits and vegetables.

Additionally the compositions can be used in drug or pharmaceuticalpreparations and cosmetics, including but not limited to toothpaste,mouthwash, cough syrup, chewable tablets, lozenges, vitaminpreparations, and the like.

The compositions can be used “as-is” or in combination with othersweeteners, flavors and food ingredients.

Non-limiting examples of sweeteners include steviol glycosides,stevioside, rebaudioside A, rebaudioside B, rebaudioside C, rebaudiosideD, rebaudioside E, rebaudioside F, dulcoside A, steviolbioside,rubusoside, as well as other steviol glycosides found in Steviarebaudiana Bertoni plant and mixtures thereof, stevia extract, Luo HanGuo extract, mogrosides, high-fructose corn syrup, corn syrup, invertsugar, fructooligosaccharides, inulin, inulooligosaccharides, couplingsugar, maltooligosaccharides, maltodextrins, corn syrup solids, glucose,maltose, sucrose, lactose, aspartame, saccharin, sucralose, sugaralcohols.

Non-limiting examples of flavors include lemon, orange, fruit, banana,grape, pear, pineapple, bitter almond, cola, cinnamon, sugar, cottoncandy, vanilla flavors.

Non-limiting examples of other food ingredients include flavors,acidulants, organic and amino acids, coloring agents, bulking agents,modified starches, gums, texturizers, preservatives, antioxidants,emulsifiers, stabilizers, thickeners, gelling agents.

The following examples illustrate preferred embodiments of theinvention. It will be understood that the invention is not limited tothe materials, proportions, conditions and procedures set forth in theexamples, which are only illustrative.

EXAMPLE 1 Preparation of Reb D Concentrated Solution

100 g of rebaudioside D produced by PureCircle Sdn Bhd (Malaysia), with98.1% purity (dry weight basis), having water solubility of 0.03% atroom temperature was mixed with 400 g of water and incubated inthermostatted oil bath. The temperature was increased at 1° C. perminute to 121° C. The mixture was maintained at 121° C. for 1 hour andthen the temperature was decreased to 80° C., at 1° C. per minute tomake Solution#1.

EXAMPLE 2 Preparation of Reb D and Reb A Concentrated Solution

70 g of rebaudioside D with 98.1% purity (dry weight basis), havingwater solubility of 0.03% and 30 g of rebaudioside A with 98.6% purity(dry weight basis), and having water solubility of 5.5%, both producedby PureCircle Sdn Bhd (Malaysia), were mixed with 400 g water andsubjected to heat treatment as described in EXAMPLE 1 to makeSolution#2.

EXAMPLE 3 Preparation of Reb D and Reb B Concentrated Solution

70 g of rebaudioside D with 98.1% purity (dry weight basis), havingwater solubility of 0.03% and 30 g of rebaudioside B with 99.0% purity(dry weight basis), and having water solubility of 0.01%, both producedby PureCircle Sdn Bhd (Malaysia), were mixed with 400g water andsubjected to heat treatment as described in EXAMPLE 1 to makeSolution#3.

EXAMPLE 4 Concentrated Solution Stability

Solution#1, solution#2 and solution#3 prepared according to EXAMPLE 1,EXAMPLE 2 and EXAMPLE 3 were assessed in terms of their stability. Theresults are summarized in Table 2.

TABLE 2 Concentrated solution stability (80° C.) Time, Observation minSolution #1 Solution #2 Solution #3 1 Cloudy solution Clear solutionClear solution 5 Intensive crystallization Cloudy solution Clearsolution 15 Viscous slurry of crystals Intensive Clear solutioncrystallization 30 Solidified crystalline Viscous slurry of Clearsolution mixture crystals 60 Solidified crystalline Solidifiedcrystalline Cloudy solution mixture mixture

It can be seen that the solution prepared by combining reb D and reb Bshows greater stability towards crystallization.

EXAMPLE 5 Preparation of Highly Soluble Rebaudioside D Compositions

Freshly prepared solution#1, solution#2 and solution#3 preparedaccording to EXAMPLE 1, EXAMPLE 2 and EXAMPLE 3 were spray dried usingYC-015 laboratory spray drier (Shanghai Pilotech Instrument & EquipmentCo. Ltd., China) operating at 175° C. inlet and 100° C. outlettemperature. Solutions were maintained at 80° C. to prevent prematurecrystallization. The solution#1 yielded sample #1, solution #2 yieldedsample #2 and solution #3 yielded sample #3.

The obtained amorphous powder samples were compared for solubility(Table 3).

TABLE 3 Solubility of Rebaudioside D compositions ObservationSolubility, % Sample #1 Sample #2 Sample #3 0.01 Clear solution Clearsolution Clear solution 0.05 Slightly cloudy Clear solution Clearsolution solution 0.5 Cloudy solution Slightly cloudy Clear solutionsolution 1.0 Visible undissolved Cloudy solution Clear solution matter1.5 Visible undissolved Visible undissolved Slightly cloudy mattermatter solution

EXAMPLE 6 Taste Profile of Rebaudioside D Compositions

Taste profiles of sample#1, sample#2 and sample#3 prepared according toEXAMPLE 5 were compared. A model zero-calorie carbonated beverageaccording to formula presented below was prepared.

Ingredients Quantity, % Cola flavor 0.340 ortho-Phosphoric acid 0.100Sodium citrate 0.310 Sodium benzoate 0.018 Citric acid 0.018 Sample #1or 2 or 3 0.050 Carbonated water to 100

The sensory properties were evaluated by 20 panelists. The results aresummarized in Table 4.

TABLE 4 Evaluation of zero-calorie carbonated beverage samples Number ofpanelists detected the attribute Taste attribute Sample #1 Sample #2Sample #3 Bitter taste 0 10 0 Astringent taste 0 12 1 Aftertaste 0 11 1Comments Quality of sweet taste Clean Bitter aftertaste Clean (20 of 20)(11 of 20) (18 of 20) Overall evaluation Satisfactory SatisfactorySatisfactory (20 of 20) (7 of 20) (17 of 20)

The above results show that the beverages prepared using thecompositions comprising Reb D and Reb B (sample#3) possessed almostsimilar taste profile with pure reb D (sample #1), at the same timeexceeding pure Reb D in solubility almost 20 times. On the other handcompositions comprising Reb D and Reb A (sample#2) possessed inferiorsolubility and taste profile compared to sample #3.

EXAMPLE 7 Preparation of Reb D and Reb B Carboxylate Salt Composition

70 g of rebaudioside D with 98.1% purity (dry weight basis), havingwater solubility of 0.03% and 30 g of rebaudioside B with 99.0% purity(dry weight basis), and having water solubility of 0.01%, both producedby PureCircle Sdn Bhd (Malaysia), were mixed with 400 g water andsubjected to heat treatment as described in EXAMPLE 1 to makeconcentrated solution. The pH of the solution was adjusted with 40% KOHto pH 6.0 and the solution was spray dried as described in EXAMPLE 5.The taste profile of obtained carboxylate salt composition was comparedwith sample#3 of EXAMPLE 5, according to procedure described in EXAMPLE6. No significant differences between sample #3 and the carboxylate saltcomposition were revealed during the comparison.

The process of the present invention resulted in a Rebaudioside Dcompositions that demonstrated high degree of solubility in water, andsuperior taste profile. Although the foregoing embodiments describe theuse of Rebaudioside D, Rebaudioside B and Rebaudioside A, it is to beunderstood that any Stevia-based sweetener may be used and prepared inaccordance with this invention, and all Stevia-based sweeteners arecontemplated to be within the scope of the present invention.

Although the invention and its advantages have been described in detail,it should be understood that various changes, substitutions andalterations can be made herein without departing from the spirit andscope of the invention as defined by the appended claims. Moreover, thescope of the application is not intended to be limited to the particularembodiments of the invention described in the specification. As one ofordinary skill in the art will readily appreciate from the disclosure ofthe invention, the compositions, processes, methods, and steps,presently existing or later to be developed that perform substantiallythe same function or achieve substantially the same result as thecorresponding embodiments described herein may be utilized according tothe invention.

We claim:
 1. A method for producing a high solubility Stevia sweetenercomprising the steps of: A) providing a first Stevia sweetener; B)providing a second Stevia sweetener that is different from the firstStevia sweetener; C) providing water; D) mixing the water and first andsecond Stevia sweeteners to make a mixture; E) increasing thetemperature of the mixture by a gradient method to make a solution; F)holding the solution at an elevated temperature; G) decreasing thetemperature of the solution by a gradient method to obtain a highstability and high concentration Stevia sweetener solution; and H) spraydrying the high stability and high concentration Stevia sweetenersolution, to provide the high solubility Stevia sweetener.
 2. The methodof claim 1 wherein the first Stevia sweetener is selected from the groupconsisting of Stevioside, Rebaudioside A, Rebaudioside B, RebaudiosideC, Rebaudioside D, Rebaudioside E, Rebaudioside F, Steviolbioside,Dulcoside A, Rubusoside, or other glycoside of steviol, and a mixturethereof.
 3. The method of claim 1 wherein the second Stevia sweetener isselected from the group consisting of: Stevioside, Rebaudioside A,Rebaudioside B, Rebaudioside C, Rebaudioside D, Rebaudioside E,Rebaudioside F, Steviolbioside, Dulcoside A, Rubusoside, or otherglycoside of steviol, and a mixture thereof.
 4. The method of claim 1wherein a ratio of the first and second Stevia sweeteners is about 99:1to 1:99, preferably about 50:50 to 95:5, more preferably about 70:30 to90:10 (w/w).
 5. The method of claim 1 wherein the first Stevia sweeteneris rebaudioside D.
 6. The method of claim 1 wherein the second Steviasweetener is rebaudioside B.
 7. The method of claim 1 wherein the secondStevia sweetener is rebaudioside A.
 8. The method of claim 1 wherein thesecond Stevia sweetener is steviolbioside.
 9. The method of claim 1wherein the second Stevia sweetener comprises a mixture of rebaudiosideA and rebaudioside B.
 10. The method of claim 1 wherein the secondStevia sweetener comprises a mixture of rebaudioside A andsteviolbioside.
 11. The method of claim 1 wherein the second Steviasweetener comprises a mixture of rebaudioside A, rebaudioside B andsteviolbioside.
 12. The method of claim 1 wherein the second Steviasweetener comprises a mixture of rebaudioside B and steviolbioside. 13.The method of claim 1 wherein a ratio of water to the combined first andsecond Stevia sweeteners is about 1:1 to 10:1, preferably about 3:1 to6:1 (v/w).
 14. The method of claim 1 wherein the water and Steviasweetener mixture is heated at a gradient of about 1° C. per minute to atemperature of about 118-125° C.
 15. The method of claim 1 wherein thewater and Stevia sweetener mixture is held at a temperature of about118-125° C. for a period of about 50-70 minutes.
 16. The method of claim1 wherein the temperature of the solution is cooled down to atemperature of about 80° C. at gradient of about 1° C. per minute toobtain the high stability and high concentration Stevia sweetenersolution.
 17. The method of claim 1 wherein high stability and highconcentration Stevia sweetener solution is spray dried on a spray dryingapparatus operating at about 175° C. inlet and about 100° C. outlettemperatures.
 18. The method of claim 1, wherein a pH of high stabilityand high concentration Stevia sweetener solution is adjusted by alkalinesolution to about pH 4.5 to 7.0, preferably about pH 5.5 to 6.5.
 19. Ahigh solubility Stevia sweetener powder made by the process of claim 1.20. The high solubility Stevia sweetener powder of claim 19, having asolubility of at least about 0.5 grams per 100 grams of water.
 21. Thehigh solubility Stevia sweetener powder of claim 19, having a solubilityof at least about 1.0 gram per 100 grams of water.
 22. The highsolubility Stevia sweetener powder of claim 19, having a solubility ofat least about 5.0 grams per 100 grams of water.
 23. The high solubilityStevia sweetener powder of claim 19, having a solubility of at leastabout 10.0 grams per 100 grams of water.
 24. A high solubility Steviasweetener made by the process of claim 1, comprising a mixture ofrebaudioside D and rebaudioside B, wherein rebaudioside B, is partiallyor fully converted to a caboxylate salt.
 25. A sweetener compositioncomprising a high solubility Stevia sweetener made by the process ofclaim 1 and an additional sweetening agent selected from the groupconsisting of: stevia extract, steviol glycosides, stevioside,rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D,rebaudioside E, rebaudioside F, dulcoside A, steviolbioside, rubusoside,other steviol glycosides found in Stevia rebaudiana Bertoni plant andmixtures thereof, Luo Han Guo extract, mogrosides, high-fructose cornsyrup, corn syrup, invert sugar, fructooligosaccharides, inulin,inulooligosaccharides, coupling sugar, maltooligosaccharides,maltodextrins, corn syrup solids, glucose, maltose, sucrose, lactose,aspartame, saccharin, sucralose, sugar alcohols, and a combinationthereof.
 26. A flavor composition comprising a high solubility Steviasweetener made by the process of claim 1 and an additional flavoringagent selected from the group consisting of: lemon, orange, fruit,banana, grape, pear, pineapple, mango, bitter almond, cola, cinnamon,sugar, cotton candy, vanilla, and a combination thereof.
 27. A foodingredient comprising a high solubility Stevia sweetener made by theprocess of claim 1 and an additional food ingredient selected from thegroup consisting of: acidulants, organic and amino acids, coloringagents, bulking agents, modified starches, gums, texturizers,preservatives, antioxidants, emulsifiers, stabilizers, thickeners,gelling agents, and a combination thereof.
 28. A food, beverage,cosmetic or pharmaceutical product comprising high solubility Steviasweetener made by the process of claim
 1. 29. A highly soluble steviacomposition comprising a mixture of rebaudioside D and rebaudioside B,at a ratio of about 50%:50% to about 80%:20% (w/w).
 30. A food,beverage, cosmetic or pharmaceutical product comprising the steviacomposition of claim 29.